Establishment Of Murine Lewis Lung Carcinoma Cell Line Which Can Express Interleukin-12 Stably And Identification Of Its Activity

Objective:Interleukin 12 (IL-12) is a proinflammatory cytokine with antitumor activity. To construct the plasmid that can express murine IL-12 gene stablely in eukaryotic cell, and establish the murine Lewis Lung Carcinoma cell(LLC) line which can express murine IL-12 gene stably through recombinant plasmid, provide basis for further study of its immunoregulatory mechanism and antitumor immune response with mIL-12.Methods:PORF- mIL-12(Elasti) plasmid was amplified through PCR ,the objective segment (mIL-12)was inserted into eukaryotic expression vector pcDNA3.1(+) after enzyme digestion with restriction enzyme Hindâ…¢and EcoRâ… , mIL-12 was drived by mCMV promoter of pcDNA3.1(+) and transcribed into the same mRNA,the recombinant plasmid was transfected into LLC cell line with lipofectin after identifying it and measured its expression, cells cycle and apoptosis was observed by FCM . Got positive clone cells after selected by G418 and cultured them largely, meantime the activity of mIL-12 was examined.Results The mIL-12 was amplified and inserted into eukaryotic expression vector pcDNA3.1(+) after enzyme digestion and interlink,...